Journal: Journal of Clinical Microbiology
Article Title: Contaminations Occurring in Fungal PCR Assays
Figure Lengend Snippet: Gel electrophoresis (2% TAE-agarose gel) showing amplified DNA of potentially contaminated samples (specific bands of approximately 500-bp length). Lanes: 1, 100-bp marker; 2, zymolyase, contaminated lot; 3, zymolyase, contaminated lot, 1:10 dilution; 4, 10× buffer from digoxigenin (Dig)-labeling kit (Boehringer Mannheim), contaminated lot; 5, 10× buffer from Dig-labeling kit (Boehringer Mannheim), contaminated lot; 6, zymolyase, regular lot, not contaminated; 7, proteinase K, regular lot, not contaminated; 8, lyticase, contaminated lot; 9, lysing enzyme extracted from T. harzianum ; 10, positive control, A. fumigatus DNA, extracted from 10 3 CFU; 11, negative control of the extraction; and 12, negative control of the PCR.
Article Snippet: Detailed and repeated serial analysis of all components of the PCR mixture (nucleotides, primers, water, magnesium chloride, 10× buffer, and Taq polymerase) showed a positive specific band only in the 10× PCR buffer supplied by Boehringer Mannheim.
Techniques: Nucleic Acid Electrophoresis, Agarose Gel Electrophoresis, Amplification, Marker, Labeling, Positive Control, Negative Control, Polymerase Chain Reaction